Use of Carophyllenes in the Manufacture of Medicaments and Treatment of Bodily Conditions of Inflammation and Inflammatory Pain

ABSTRACT

The invention concerns the use of caryophyllenes related to medicaments and to the treatment of bodily conditions of inflammation and inflammatory pain.

FIELD OF THE INVENTION

The present invention concerns the use of caryophyllenes related tomedicaments and to the treatment of bodily conditions of inflammationand inflammatory pain. It relates particularly to the use ofcaryophyllenes in the manufacture of medicaments for the treatment ofinflammatory conditions of the animal body, including the human body.The invention also concerns the use of caryophyllenes for the treatmentof inflammatory conditions of the body, including inflammatory pain.

BACKGROUND OF THE INVENTION

Caryophyllenes are known chemical compounds, useful in variousapplications. For instance patent document U.S. Pat. No. 3,987,008reveals sesquiterpenic derivatives as odor and taste-modifying agents;In J. Nat. Prod. 1992 July; 55(7):999-1003, beta-caryophyllene andalpha-humulene as cited as potential anticarcinogenic agents; in patentapplication WO alpha and beta-humulene and (−)-beta-caryophyllene arecited in the control of whitefly species; in U.S. Pat. No. 5,314,693alpha-humulene is cited as a repellent for pine wood nematodes; inpatent application WO 02078719 alpha and beta-caryophyllene arecomprised in antitumor compositions.

SUMMARY OF THE INVENTION

The present invention concerns, in one particular aspect, a new anduseful use for caryophyllenes, more particularly alpha-humulene orbeta-caryophyllene, as anti-inflammatory and as analgesic, in a broadsense. In a more specific sense, the compounds were found to be usefulinhibitors of entities that are known to be involved in the inflammatoryprocess:

pro-inflammatory cytokines IL-1β (interleukin 1β) and TNFα (tumornecrosis factor α);

PGE2 (prostaglandin-E2),

Expression of COX-2 (cycloxigenase-2) and iNOS (inducible nitric oxidesynthase) enzymes

One particular example of caryophyllene of the invention isalpha-humulene, a sesquiterpene identified with the CAS (ChemicalAbstracts Service) registry number 6753-98-6, also known asalpha-caryophyllene, represented by the following structure:

Another particular example of caryophyllene of the invention is thetrans-caryophyllene (or beta-caryophyllene), also a sesquiterpene,identified with the CAS (Chemical Abstracts Service) registry number87-44-5, represented by the following alternative structures A and B:

According to the meaning employed herein, mention to caryophyllenes,object of the invention, includes the molecules as such, their salts,isomers, metabolites, pro-drugs, solvates (including hydrates) andadducts.

Terpenes, including sesquiterpenes, are often mentioned as components ofcomplex mixtures extracted from plants, where—as known to a personsskilled in the art—it is undetermined what compound or compounds areeffective, how much effective they are, and whether they are active bythemselves, by way of the vehicle/solvent the composition contains(water, alcohol, other solvents, mixtures of those, etc), or by way oftheir interaction with other components within the mixtures. Individualterpenes per se, of natural origin or products of synthesis (forinstance J. Am. Chem. Soc., 99, 3864 (1977)), are rarely mentioned aseffective pharmaceutical agents. Alpha-humulene has even been mentionedto be virtually inactive concerning anti-inflammatory or chemo-therapiceffects (reference: Carcinogenesis (2002), 23(5), 795-802).

The applicant has now found out that caryophyllenes, particularlyalpha-humulene and trans-caryophyllene, have marked anti-inflammatoryeffects, including inflammatory pain, comprised therein the inhibitoryeffect upon the production of pro-inflammatory cytokines IL-1β and TNFα,prostaglandin PGE2, or the expression of enzymes COX-2 and iNOS.

The applicant has also found out that caryophyllenes, particularlyalpha-humulene and trans-caryophyllene, have anti-allergic, particularlyanti-histaminic effects.

The caryophyllenes of the invention are part of the ongoing search fordrugs with direct of indirect inflammatory activity, which inhibit thephysiopathology processes involved in inflammation. They are used in thecontrol of chronic-degenerative diseases as rheumatoid arthritis,osteoarthritis, systemic lupus eritematosus, ulcerative colitis,psoriasis, atopic eczema, atherosclerosis, and other non-degenerativediseases as depression, and cellulites, and allergies.

Therefore, one of the objects of the present invention is the use ofcaryophyllenes, particularly alpha-humulene and/or trans-caryophyllene,or compositions comprising caryophyllenes, in the manufacture of amedicament for the treatment of inflammatory conditions of the animalbody, particularly the human body.

Another object of the present invention is the use of caryophyllenes,particularly alpha-humulene and trans-caryophyllene, or compositionscontaining caryophyllenes, in the treatment of inflammatory conditionsof the animal body, particularly the human body.

Another object of the present invention is a method of treatment of aninflammatory condition of the animal body, particularly the human body,comprising the administration of a therapeutically effective amount ofcaryophyllenes, particularly alpha-humulene or trans-caryophyllene, to apatient.

Another object of the present invention is the use of caryophyllenes,particularly alpha-humulene and trans-caryophyllene, or compositionscontaining caryophyllenes, for the inhibition of the bodily productionof one or more of cytokine IL-1β, cytokine TNFα, prostaglandin PGE2,expression of enzymes COX-2 and iNOS.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

The caryophyllene of the invention, as well as compositions comprisingthe caryophyllene according to the invention, can be administered to thesubject in need of treatment in any adequate way, enteral or parenteral,including oral, topical, transdermal, subcutaneous, intraperitonial,intravenous, by infiltration, by inhalation, transdermal, transmucosal,intramuscular, intrapulmonary, vaginal, rectal, intraocular, andsublingual. Particularly adequate ways of administration in the presentinvention are systemically (infiltration, oral, inhalation by spray,transdermal) and topically. The caryophyllene of the invention can becomprised in a slow or controlled release composition. Known adjuvantsand excipients can be utilized in the compositions. A reference forpharmaceutical dosage forms useful for the compositions related to theinventions can be found in the publication Remington's PharmaceuticalSciences, Mack Publishing.

The compositions comprising caryophyllene can be administered topatients as solids, liquids or semi-liquids, tablets, capsules, pills,powder, granules, suspensions, emulsions, dispersions and any otheruseful known form.

The compositions might contain further active agents, for instanceantibiotics, depending on the desired effect.

For oral administration as tablets or capsules (both soft and hardcapsules), the caryophyllene can be combined with pharmaceuticallyacceptable inert vehicles, such as lactose, starch, sucrose, glucose,methyl cellulose, magnesium stearate, dicalcium phosphate, calciumphosphate, manitol, sorbitol, and similars; for oral administration inthe liquid form, the caryophyllenes can be combined with ethanol,glycerol, water, and similars. When desired or necessary, agglomeratingagents, lubricant agents, disintegrating agents, color and fragrance canbe added to the mixture. Common agglomerating agents are glucose,β-lactose, corn sweeteners, natural or synthetic gums such as gumarabica, tragacanth or sodium alginate, carboxymethylcellulose,polyethylene glycol, wax and similars. Lubricants include sodium oleate,sodium stearate, magnesium stearate, sodium benzoate, sodium acetate,sodium chloride. Disintegrants include starch, methyl cellulose, agar,bentonite, xanthan gum, and similars.

The compositions concerned in the invention can also be administrated asliposomes or coupled with soluble polymers as vehicles.

Liquid dosage forms for oral administration may comprise colorants andedulcorants to increase acceptance by patients. Acceptable vehicles forwater dosage forms are, water, an appropriate oil, a saline solution,aqueous dextrose, other sugar solutions and glycols as propylene glycolor polyethylene glycols, phosphate buffer.

Compositions related to the present invention typically comprise about 1mg to about 1000 mg of one or more caryophyllenes, particularly about 10to 200 mg and more particularly about 30 to 100 mg. In such compositionsthe caryophyllene represents about 0.1 to 99% in weight, particularlyabout 1 to 70% and more particularly about 10 to 40%, optionallycomprising at least one pharmaceutically acceptable vehicle.

EXAMPLES

The examples that follow represent particular embodiments of theinvention, and do not impose any limitation to its extension, which islimited only by the claims attached hereto.

Example 1 Inflammatory Nociception Induced by Carregenin

The evaluation methodology used in this test is described by Vaz et al.in J. Pharmacol. Exp. Ther. 278:304-312, 1996.

Male mice (25-35 g) were systemically (orally) treated withalpha-humulene, 50 mg/kg, administered 1 hour before the experiment.Animals treated with 0.9% saline solution (0.1 ml/10 g) were used as thecontrol. Another group of animals was treated with paracetamol (600mg/kg, orally, administered 1 h before the treatment), that was used aspositive control. For the induction of inflammatory pain, the animalsreceived an intraplantar injection of 0.05 ml of carrageenin (300 μg perpaw) at the plantar surface of the right hind paw. This dosage causesoedema, nociception and substantial swelling of the injected paw.

The nociception was evaluated with a Von Frey filament (0.4 g) after 3,4 and 6 hours. To obtain a basal response, the animals were pre-testedthe previous day with the 0.4 g von Frey filament. Only animals with aresponse of about 20% were selected. The filament was applied to theright hind paw, complying with the criteria of (1) the application wasperpendicular to the plantar surface, with enough pressure to cause thefilament to bend, thus obtaining total pressure; (2) the animals wereevaluated when the four paws were touching the screen; (3) the pawwithdrawal response was considered when the animal removed the pawentirely from the support screen; (4) each animal was stimulated 10consecutive times, each stimulation lasting 1 second; (5) each pawwithdrawal event was considered as 10% of the response, with 10withdrawal events corresponding to 100% response.

Graph 1 below compares the pain inhibition obtained by alpha-humulenewith the administration of paracetamol. Each point represents theaverage of 5 animals, and the vertical bars the mean standard errordeviation.

The graph clearly shows that, according to the invention, acaryophyllene as alpha-humulene reduced the inflammatory nociception, asa result of reduction of inflammation, as much as a known analgesic,paracetamol.

Example 2 Carrageenin Oedema in Mouse Paw

The test used below is described by Cunha et. al. in the publicationLife Sci. 70:159-169, 2001.

Male 25 g-35 g mice were slightly sedated with ether and were injected50 μl saline containing carrageenin (300 μl/paw) in the right paw. Theleft paw received the same volume of saline and was taken as a negativecontrol. The swelling was measured with a plethysmometer (manufacturer:Ugo Basile, Italy) along various time intervals after the injection ofthe phlogistic agent. The difference between the volumes of the rightand the left paw were quantified (in ml) and taken as an index ofoedema. One hour before the test the animals were systemically treated(orally) with 50 mg/kg of alphahumulene or trans-caryophyllene.

Graph 2 below compares the inhibition of the volume of the oedema byadministration of either alpha-humulene or trans-caryophyllene withinhibition obtained with the administration of dexamethasone (0.5 mg/kg,injected subcutaneously 4 h before test) and used as positive control.Oedema volume measurement time point intervals were 30, 60, 120 and 240min, 24 h and 48 h. Each point represents the average of 5 animals, andthe vertical bars the mean standard error deviation.

The graph clearly shows that, according to the invention, acaryophyllene as alpha-humulene reduced the inflammatory volume, as diddexamethasone.

Example 3 Bradykinin Oedema in Mouse Paw

The test used below is described by Cunha et. al. in the publicationLife Sci. 70:159-169, 2001.

Male 25 g-35 g mice were slightly sedated with ether and were injected50 μl saline containing bradykinin (BK, 3 nmol/paw), intraplantar, inthe right paw. The left paw received the same volume of saline and wastaken as negative control. The swelling was measured with aplethysmometer (manufacturer: Ugo Basile, Italy) along various timepoints intervals after the injection of the phlogistic agent. Thedifference between the volumes of the right and the left paw werequantified (in ml) and taken as an index of oedema. One hour before thetest the animals were systemically treated (orally) with 50 mg/kg ofalpha-humulene or with trans-caryophyllene.

The animals were pre-treated with 5 mg/kg of captopril, injectedsubcutaneously, 1 hour before the test, in order to avoid degradation ofkinines.

Graphs 3A and 3B below compare the inhibition of the volume of theoedema by administration of alpha-humulene (3A) or trans-caryophyllene(3B). Oedema volume measurement time intervals were 10, 20, 30, 60, and120 min, 24 h and 48 h. Each point represents the average of 5 animals,and the vertical bars the mean standard error deviation.

The graph clearly shows that, according to the invention, acaryophyllene as alpha-humulene or trans-caryophyllene markedly reducedbradykinin-induced paw oedema.

Example 4 Histamine Oedema in Mouse Paw

The test used below is described by Cunha et. al. in the publicationLife Sci. 70:159-169, 2001.

Male 25 g-35 g mice were slightly sedated with ether and were injected50 μl saline containing histamine (100 nmol/paw), intraplantar, in theright paw. The left paw received the same volume of saline and was takenas negative control. The swelling was measured with a plethysmometer(manufacturer: Ugo Basile, Italy) along various time intervals after theinjection of the phlogistic agent. The difference between the volumes ofthe right and the left paw were quantified (in ml) and taken as an indexof oedema. One hour before the test the animals were systemicallytreated (orally) with 50 mg/kg of alpha-humulene.

Graph 4 below compares the inhibition of the volume of the oedema byadministration of alpha-humulene. Oedema volume measurement timeintervals were 10, 20, 30, 60, and 120 min, 24 h and 48 h. Each pointrepresents the average of 5 animals, and the vertical bars the meanstandard deviation.

The graph clearly shows that, according to the invention, acaryophyllene as alpha-humulene significantly reduced histamine-inducedoedema formation. It also indirectly shows effect against allergy.

Example 5 Platelet Aggregation Factor (PAF) Oedema in Mouse Paw

The test used below is described by Cunha et. al. in the publicationLife Sci. 70:159-169, 2001.

Male 25 g-35 g mice were slightly sedated with ether and were injected50 μl saline containing platelet aggregation factor (PAF, 3 nmol/paw),intraplantar, in the right paw. The left paw received the same volume ofsaline and was taken as negative control. The swelling was measured witha plethysmometer (manufacturer: Ugo Basile, Italy) along various timeintervals after the injection of the phlogistic agent. The differencebetween the volumes of the right and the left paw were quantified (inml) and taken as an index of oedema. One hour before the test theanimals were systemically treated (orally) with 50 mg/kg ofalpha-humulene or trans-caryophyllene.

Graph 5 below compares the inhibition of the volume of the oedema byadministration of alpha-humulene (SA) and trans-caryophyllene (5B).Oedema volume measurement time intervals were 30, 45, 60, and 120 min.Each point represents the average of 5 animals, and the vertical barsthe mean standard deviation.

The graph clearly shows that, according to the invention, acaryophyllene as alpha-humulene or trans-caryophyllene markedly reducedPAF-induced oedema formation. As PAF is also known to be involved inallergic processes, such data further reinforces the use ofcaryophyllenes in the management of allergic states.

Example 6 Arachidonic Acid Oedema in Mouse Ear

The ear oedema in the test below was measured according to Calixto et.al. in the publication Prostaglandins, 5: 515-526, 1991, with minormodifications.

Male 25 g-35 g mice, in a first group, were topically applied, in theinner surface of the ears, an ointment comprising a range from 0.025 to0.2% alpha-humulene or trans-caryophyllene. In the positive controlgroup, the animals were topically applied 0.05 mg of phenidone per ear.After 60 minutes, the animals received 20 μl of arachidonic acid (2mg/ear), dissolved in acetone, in the inner surface of the right ear.The oedema was measured using a digital micrometer, and the responseswere expressed as μm, the difference between the ear thickness beforeand after the application of arachidonic acid. The responses of theanimals treated with caryophyllenes were compared to those observed inthe control group animals, treated with base ointment.

Graphs 6A and 6B below compare the inhibition of the volume of theoedema by topic administration of alpha-humulene (6A) andtrans-caryophyllene (6B). Oedema volume measurement was performed afterapplication of 0.025%, 0.05%, 0.1, and 0.2% caryophyllene contentointment, compared to the oedema volume caused by the application ofphenidone and arachidonic acid (C). Each point represents the average of5 animals, and the vertical bars the mean standard error deviation.

The graphs clearly show that, according to the invention, the topicapplication of a caryophyllene as alpha-humulene or trans-caryophyllenemarkedly reduced oedema formation, in a dose-dependent manner.

Example 7 Levels of Pro-Inflammatory Cytokine IL-1B

The test used below is described by Campos et. al. in the publicationBr. J. Pharmacol. 135: 1107-1114, 2002, with minor modifications.

Male 160-180 g rats were orally given 50 mg/kg of alpha-humulene.Animals treated with 0.9% (0.1 ml/10 g) saline were used as control.Another group of animals was treated with 0.5 mg/kg dexamethasone,subcutaneously, 4 hours before the test, and used as positive control.After 60 minutes, the animals received intraplantar injections of 100 μlof carrageenin (300 μg/paw) and were sacrificed after 180 minutes.Control animals received saline. The subcutaneous tissue of the injectedpaws was removed and put in a phosphate buffer containing 0.5% tween 20,0.1 mM benzametonium chloride, 10 mM EDTA, 2 μg/m aprotinin, 0.1 mM PMSF(phenyl methyl sulfonyl fluoride) and 0.5% BSA (bovine serum albumin).The tissues were homogenized and centrifuged at 3000 g, for 10 min, at−4° C. The supernatant was used in the test. The levels of IL-1β weremeasures with an Elisa kit, according to the manufacturer's instructions(R & D Systems®, USA). The tests were performed in duplicate, andrepeated three times. The answers are expressed in pg/mg of tissue.

Graph 7 below (each result represents the average of 5 animals, and thevertical bars the mean standard error deviation) compares the inhibitionof production of inflammatory cytokine IL-1β induced by carrageenin inthe paws of rats.

The graph clearly shows that, according to the invention, theadministration of a caryophyllene, such as alpha-humulene, markedlyinhibited the production of pro-inflammatory cytokine IL-1β induced bycarrageenin in the paws of rats.

Example 8 Levels of Pro-Inflammatory Cytokine TNFα

The test used below is described by Campos et. al. in the publicationBr. J. Pharmacol. 135: 1107-1114, 2002, with minor modifications.

Male 60-180 g rats were orally given 50 mg/kg of trans-caryophyllene.Animals treated with 0.9% (0.1 ml/10 g) saline were used as control.Another group of animals was treated with 0.5 mg/kg dexamethasone,subcutaneously, 4 hours before the test, and used as positive control.After 60 minutes, the animals received intraplantar injections of 100 μlof carrageenin (300 μg/paw) and were sacrificed after 180 minutes.Control animals received saline. The subcutaneous tissue of the injectedpaws was removed and put in a phosphate buffer containing 0.5% tween 20,0.1 mM benzametonium chloride, 10 mM EDTA, 2 μg/m aprotinin, 0.1 mM PMSF(phenyl methyl sulfonyl fluoride) and 0.5% BSA (bovine serum albumin).The tissues were homogenized and centrifuged at 3000 g, for 10 min, at−4° C. The supernatant was used in the test. The levels of TNFαweremeasures with an Elisa kit, according to the manufacturer's instructions(R & D Systems®, USA). The tests were performed in duplicate, andrepeated three times. The answers are expressed in pg/mg of tissue.

Graph 8 below (each result represents the average of 5 animals, and thevertical bars the mean standard error deviation) compares the inhibitionof production of inflammatory cytokine TNFα induced by carrageenin inthe paws of rats.

The graph clearly shows that, according to the invention, theadministration of a caryophyllene, such as trans-caryophyllene, markedlyinhibited the production of the pro-inflammatory cytokine TNFα inducedby carrageenin in the paws of rats.

Example 9 Levels of PGE2

The test used below is described by Pinheiro et. al. in the publicationInflamm. Res. 51: 603-610, 2002, with minor modifications.

Male 160-180 g rats were orally given 50 mg/kg of trans-caryophyllene.Animals treated with 0.9% (0.1 ml/10 g) saline were used as control.Another group of animals was treated with 0.5 mg/kg dexamethasone,subcutaneously, 4 hours before the test, and used as positive control.After 60 minutes, the animals received intraplantar injections of 100 μlof carrageenin (300 μg/paw) and were sacrificed after 180 minutes.

The exsudate of the paws was collected by dialysis with the help of twopolyethylene canulas, and was utilized for the quantification of PGE2,with an Elisa kit, according to the manufacturer's instructions (R & DSystems®, USA). The tests were performed in duplicate, and repeatedthree times. The answers are expressed in pg/mg of tissue.

Graph 9 below (each result represents the average of 5 animals, and thevertical bars the mean standard error deviation) compares inhibition ofthe PGE2 level growth induced by carrageenin in the paws of rats, byadministration of alpha-humulene and trans-caryophyllene, compared tothe effect obtained by treatment with dexomethasone.

The graph clearly shows that, according to the invention, theadministration of a caryophyllene, such as alpha humulene ortrans-caryophyllene, markedly inhibited the growth of PGE2 levelsinduced by carrageenin in the paws of rats.

Example 10 Inhibition of the Expression of Enzymes Cox-2 and iNOS

The expression of enzymes COX-2 and iNOS were determined by Western blotaccording to the methodology described by Medeiros et. al. in thepublication Circ Res. 28:1375-1382, 2004.

Male 160-180 g rats were orally given 50 mg/kg of trans-caryophyllene.Animals treated with 0.9% (0.1 ml/10 g) saline were used as control.Another group of animals was treated with 0.5 mg/kg dexamethasone,subcutaneously, 4 hours before the test, and used as positive control.After 60 minutes, the animals received intraplantar injections of 100 μlof carrageenin (300 μg/paw) and were sacrificed after 180 minutes, andsubcutaneous paw tissue was removed 240 minutes after the carrageenininjection.

The collected tissue was immediately frozen in liquid nitrogen andre-suspended in a buffer of hypotonic lysis (10 mM HEPESN-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid, 1.5 mM MgCl2, 10 mMKCl, 0.5 mM PMSF phenylmethylsulphonyl fluoride, 1.5 μg/ml trypsininhibitor, 7 μg/ml pepstatin A, 5 μg/ml leupeptin, 0.1 mM benzamidine0.1 mM and 0.5 mM dithiothreitol) and homogenized. The homogenate wasdivided in three 2 ml aliquots, cooled in ice for 15 minutes, vigorouslyagitated and once again cooled in ice, in the presence of 20 μl 10%non-ionic detergent Nonidet P-40 (Roche Diagnostics, USA). The nuclearfraction was precipitated by centrifugation (1,500 g, 5 minutes) and thesupernatant containing the cytosolic extract was stored at −70° C. forthe Western blot tests. The protein concentration was determined by theBradford method (BioRad Laboratories Inc. kit, Milan, Italy). Theextracts were boiled with v/v equivalent amounts of Laemmly buffer (125mM of Tris-HCl, 2 mM of EDTA, 4% of dodecyl sodium sulphate, 20% ofglycerol, 10% of 2-mercaptoethanol and 0.1% of Comassie brilliant blue,pH 6.8). The proteins were transferred to nitrocellulose membranes (100μg/well) and separated by electrophoresis. The membranes were laterblocked by overnight incubation (4° C.) with skimmed powder milk (10%PBS), and then incubated with the anti-iNOS or anti-COX-2 antibodies for1 h at room temperature. The membranes were washed three times with 10%Triton-X in PBS with the antibody peroxidase conjugated (anti-rabbit).The bands thus obtained were quantified using a chemoluminescence kitand densitometry analysis (relative units) in radiographic films.

Graph 10 below (each result represents the average of 5 animals, and thevertical bars the mean standard error deviation) compares inhibition ofexpression of the COX2 enzymes, obtained by the administration ofalpha-humulene and trans-caryophyllene, when evaluated in thesubcutaneous tissue or the paw injected with carrageenin, compared tothe expression of COX2 induced by carrageenin obtained by treatment withdexomethasone.

The graph clearly shows that, according to the invention, theadministration of a caryophyllene, such as alpha humulene ortrans-caryophyllene, markedly inhibited the expression of enzymes COX2induced by carrageenin in the paws of rats.

Graph 11 below (each result represents the average of 5 animals, and thevertical bars the mean standard error deviation) compares inhibition ofexpression of the iNOS enzymes, obtained by the administration ofalpha-humulene, when evaluated in the subcutaneous tissue or the pawinjected with carrageenin, compared to the expression of iNOS induced bycarrageenin obtained by treatment with dexomethasone.

The graph clearly shows that, according to the invention, theadministration of a caryophyllene, such as alpha humulene, markedlyinhibited the expression of enzymes iNOS induced by carrageenin in thepaws of rats.

The examples and the information provided herein concern particularembodiments of the present invention, which is only limited by thebreath of the claims attached hereto.

1. Use of caryophyllenes, or compositions comprising caryophyllenes,characterized by the fact that it is in the manufacture of a medicamentfor the treatment of inflammatory conditions and inflammatory pain ofthe animal body, particularly the human body.
 2. Use of caryophyllenes,or compositions comprising caryophyllenes, characterized by the factthat it is in the treatment of inflammatory conditions and inflammatorypain of the animal body, particularly the human body.
 3. Use ofcaryophyllenes, or compositions comprising caryophyllenes, characterizedby the fact that it is for the inhibition of the bodily production ofone or more of the group comprising cytokine IL-1β, cytokine TNFα,prostaglandin PGE2, or expression of enzymes COX-2 and iNOS.
 4. Use ofcaryophyllenes, or compositions comprising caryophyllenes, characterizedby the fact that it is in the manufacture of a medicament for theinhibition of the bodily production of one or more of the groupcomprising cytokine IL-1β, cytokine TNFα, prostaglandin PGE2, orexpression of enzymes COX-2 and iNOS.
 5. Use of caryophyllenes accordingto one of claims 1-4 characterized by the fact that is in the treatmentof chronic-degenerative diseases comprised in the group of rheumatoidarthritis, osteoarthritis, systemic lupus eritematosus, ulcerativecolitis, psoriasis, atopic eczema, atherosclerosis, or in the treatmentof non degenerative diseases comprised in the group of depression, andcellulites, and allergies.
 6. Use of caryophyllenes according to one ofclaims 1-4 characterized by the fact that it is in the manufacture of amedicament for the treatment of chronic-degenerative diseases comprisedin the group of rheumatoid arthritis, osteoarthritis, systemic lupuseritematosus, ulcerative colitis, psoriasis, atopic eczema,atherosclerosis, or in the treatment of non degenerative diseasescomprised in the group of depression, and cellulites, or allergies. 7.Use of caryophyllenes, or compositions comprising caryophyllenes,according to one of claims 1 to 4, characterized by the fact that it isadministered via enteral or parenteral, including oral, topical,transdermal, subcutaneous, intraperitonial, intravenous, byinfiltration, by inhalation, transdermal, transmucosal, intramuscular,intrapulmonary, vaginal, rectal, intraocular, and sublingual.
 8. Useaccording to claim 7, characterized that said administration is topicalor systemical, particularly chosen among infiltration, oral, inhalationor transdermal.
 9. Use of caryophyllenes, or compositions comprisingcaryophyllenes, according to one of claims 1 to 4 characterized by thefact that said caryophyllenes are one or more of alpha-humulene andtrans-caryophyllene.
 10. A method of treatment of inflammatoryconditions and inflammatory pain of the animal body, particularly thehuman body, characterized by the fact that it comprises theadministration of a therapeutically effective amount of caryophyllenes,to a patient.
 11. A method according to claim 10 characterized by thefact that said inflammatory conditions and inflammatory pain are presentin chronic-degenerative diseases comprised in the group of rheumatoidarthritis, osteoarthritis, systemic lupus eritematosus, ulcerativecolitis, psoriasis, atopic eczema, atherosclerosis, or in the nondegenerative diseases comprised in the group of depression, andcellulites, or in allergies.
 12. A method of inhibiting the bodilyproduction of one or more of cytokine IL-1β, cytokine TNFα,prostaglandin PGE2, or expression of enzymes COX-2 and iNOS,characterized by the fact that it comprises the administration of atherapeutically effective amount of caryophyllenes, to a patient.
 13. Amethod according to one of claims 10 to 12 characterized by the factthat said caryophyllene is one or more of alpha-humulene andtrans-caryophyllene.
 14. A composition comprising caryophyllenecharacterized by the fact that the amount of said caryophyllene is about1 to about 1000 mg.
 15. A composition according to claim 14characterized by the fact that said amount is about 10 to about 200 mg.16. A composition according to claim 14 characterized by the fact thatsaid amount is about 30 to about 100 mg.
 17. A composition according toclaim 14 characterized by the fact that said caryophyllene comprisesabout 0.1 to about 99% in weight of said composition.
 18. A compositionaccording to claim 14 characterized by the fact that said caryophyllenecomprises about 1 to about 70% in weight of said composition.
 19. Acomposition according to claim 14 characterized by the fact that saidcaryophyllene comprises about 10 to about 40% in weight of saidcomposition.
 20. A composition according to one of claims 15 to 19characterized by the fact that said caryophyllene is one or more ofalpha-humulene and trans-caryophyllene.